Background: Probiotic supplements have shown the potential to reduce gastrointestinal (GI) distress symptoms; however, there is currently little research on dietary probiotic intake or the impact of probiotic food consumption on GI symptoms, particularly among athletes. Purpose: The purpose of this project was to: 1) develop and evaluate a tool for assessing intake of probiotic foods among collegiate athletes, 2) assess intake of probiotic and fermented foods among collegiate athletes at Virginia Tech, and 3) explore associations between intake of these items and GI symptoms in this study population. Methods: A literature review, review of 360 dietary recalls previously collected from collegiate athletes (n=120), and evaluation by five experts in sports nutrition and/or probiotic foods informed the development of a brief probiotic food frequency questionnaire (PRO-Q). The PRO-Q was then administered to participants (n=42), all of whom were members of Division I sports teams at Virginia Tech. Validity was assessed by comparing PRO-Q responses with probiotic intake determined using three 24-hour diet recalls (reference method). Participants also completed a questionnaire to evaluate type and severity of GI symptoms. Results: Overall intake of probiotic/fermented foods in this population was low (0.77 servings/day), with only 17% of participants reporting daily consumption. Based on Spearman’s Rho (r=0.766, p<0.001), Wilcoxon Signed-Rank test (p=0.168, indicating no difference between methods), and Bland-Altman analyses (95% agreement), PRO-Q responses were consistent with dietary recalls for average daily intake of probiotic/fermented foods. A total of 52% of participants reported experiencing regular GI distress symptoms. There was no association between PRO-Q responses and overall GI symptom score (r = 0.297, p = 0.056). Conclusion: These findings suggest that among collegiate athletes, probiotic/fermented food intake was generally low and GI distress symptoms were prevalent. The PRO-Q shows promise as a brief, self-administered method to assess probiotic/fermented food intake among collegiate athletes, although additional research is needed to confirm reproducibility and generalizability in larger samples.